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Isolation and characterization of a sperm-specific gene family in the nematode Caenorhabditis elegans.

机译:线虫秀丽隐杆线虫中精子特异基因家族的分离和鉴定。

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The major sperm protein (MSP) of the nematode Caenorhabditis elegans is a low-molecular-weight (15,000) basic protein implicated in the pseudopodial movement of mature spermatozoa. Its synthesis occurs in a specific region of the gonad and is regulated at the level of transcription (M. Klass and D. Hirsh, Dev. Biol. 84:299-312, 1981; S. Ward and M. Klass, Dev. Biol. 92:203-208, 1982; Klass et al., Dev. Biol. 93:152-164, 1982). A developmentally regulated gene family has been identified that codes for this MSP. Whole genomic blots, as well as analysis of genomic clone banks, indicate that there are between 15 and 25 copies of the MSP gene in the nematode genome. Southern blot analysis also indicates that there is no rearrangement or amplification within the MSP gene family during development. No evidence was found of methylation at various restriction sites surrounding the MSP gene family, and similarly, no correlation between methylation and expression was observed. Three distinct members of this MSP gene family have been cloned, and their nucleotide sequences have been determined. Differential screening of a cDNA clone bank made from polyadenylated mRNA from adult males yielded 45 male-specific clones, 32 of which were clones of MSP genes. One of these cDNA clones was found to contain the entire nucleotide sequence for the MSP, including part of the 5' leader and all of the 3' trailing sequence. Genomic clones bearing copies of the MSP genes have been isolated. At least one of the members of this gene family is a pseudogene. Another member of the MSP gene family that has been cloned from genomic DNA contains the entire uninterrupted structural sequence for the MSP in addition to a 5' flanking sequence containing a promoter-like region with the classic TATA box, a sequence resembling the CAAT box, and a putative ribosome-binding sequence. The 3' trailing sequences of the genomic and the cDNA clones contain an AATAAA polyadenylation site.
机译:线虫秀丽隐杆线虫的主要精子蛋白(MSP)是一种低分子量(15,000)的碱性蛋白,与成熟精子的伪足运动有关。它的合成发生在性腺的特定区域,并在转录水平上受到调节(M. Klass和D. Hirsh,Dev。Biol。84:299-312,1981; S。Ward和M. Klass,Dev。Biol 92:203-208,1982; Klass等,Dev.Biol.93:152-164,1982)。已经鉴定出发育调控的基因家族,其编码该MSP。整个基因组印迹以及对基因组克隆库的分析表明,线虫基因组中有15至25个MSP基因拷贝。 Southern印迹分析还表明,在发育过程中,MSP基因家族内没有重排或扩增。在MSP基因家族周围的各种限制性酶切位点均未发现甲基化的证据,类似地,未观察到甲基化与表达之间的相关性。已克隆了该MSP基因家族的三个不同成员,并确定了其核苷酸序列。差异筛选由成年雄性的多腺苷酸化mRNA制成的cDNA克隆库,产生了45个雄性特异性克隆,其中32个是MSP基因克隆。发现这些cDNA克隆之一包含MSP的完整核苷酸序列,包括5'前导序列的一部分和所有3'尾随序列。已经分离出带有MSP基因拷贝的基因组克隆。该基因家族的至少一个成员是假基因。从基因组DNA中克隆出来的MSP基因家族的另一个成员,除了包含一个带有经典TATA框(类似于CAAT框)的启动子样区域的5'侧翼序列之外,还包含MSP的整个不间断结构序列,和假定的核糖体结合序列。基因组和cDNA克隆的3'尾部序列含有AATAAA聚腺苷酸化位点。

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